Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
  • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
  • C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
  • C07D311/06—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2
  • C07D311/08—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 not hydrogenated in the hetero ring
  • C07D311/16—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 not hydrogenated in the hetero ring substituted in position 7
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P3/00—Drugs for disorders of the metabolism
  • A61P3/06—Antihyperlipidemics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P37/00—Drugs for immunological or allergic disorders
  • A61P37/08—Antiallergic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P39/00—General protective or antinoxious agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
  • A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system
  • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system
  • A61P9/12—Antihypertensives
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
  • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
  • C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
  • C07D311/06—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2
  • C07D311/08—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 not hydrogenated in the hetero ring
  • C07D311/18—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 not hydrogenated in the hetero ring substituted otherwise than in position 3 or 7
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
  • C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
  • C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms
  • C07H17/06—Benzopyran radicals
  • C07H17/065—Benzo[b]pyrans
  • C07H17/075—Benzo[b]pyran-2-ones

Definitions

• the present invention relates to novel coumarin derivatives of formula (I):
• n zero, 1, 2, 3 or4;
• R 5 and R 6 which can be the same or different, are optionally unsaturated C 1 -C 4 alkyl groups, or together with the nitrogen atom they form a residue of cyclic amines optionally containing other heteroatoms;
• R 1 ⁇ CH 3 or phenyl
• R 2 and R 4 which can be the same or different, are H, OH, allyl, halogen or methyl;
• R 3 ⁇ H, straight or branched, saturated or unsaturated C 1 -C 10 alkyl, which can bear OH groups, amido groups, residues of simple or derivatized sugars, or residues of optionally derivatized amino acids, or can be an optionally branched alkylene chain, (“spacer”), which links together the two residues, which can be the same or different, of formula (II)
• the invention also comprises the salts of said compounds, particularly those with pharmaceutically acceptable bases or acids, and the processes for the preparation thereof.
• novel coumarin derivatives and the salts according to the present invention have interesting pharmacological properties and may therefore be used to advantage to treat major pathologies such as peripheral ischaemia and organ ischaemia, electrical alterations of the myocardium and other organs resulting from the release of pro-inflammatory molecules (TNF, IL-1, NO, etc), peripheral and cerebral vasculopathies, angina-type disorders, hypercholesterolaemia, systemic infections such as sepsis, allergic pathologies such as asthma, rhinitis, eczema, dermatitis, such as antithrombotics and antihypertensives.
• the present invention also includes pharmaceutical preparations containing one or more of said derivatives or their salts in the form of capsules, tablets, injectable solutions, sprays and controlled release systems, creams, gels and transdermal systems.
• Coumarins include a large class of phenol substances that are to be found in plants, and are constituted by a benzene ring and an ⁇ -pyrone ring fused together. To date, at least 1,300 coumarins have been identified, mainly as metabolites of green plants, in fungi and in bacteria.
• Cloricromene (commercial name Proendotel) belongs to a family of coumarins and is prepared by the process described in U.S. Pat. Nos. 4,296,039 and 4,452,811.
• Cloricromene also has a documented effect in models of ischaemia and reperfusion in various organs, in which inflammatory-type cytokines are also involved, such as TNF (Squadrito et al.: Life Sciences 1993, 53, 341-355), and its activity is also known in the treatment of pathologies linked with vasodilatory processes and tissue damage characterised by an increased production of nitrogen oxide (patent by the Applicant No.
• IT 1265665 such as, for example, pulmonary inflammation, oedema, erythema, dermatitis, psoriasis, skin ulcers, arthritis, rheumatoid arthritis and other autoimmune disorders, hypotensive shock, septic shock, hypovolemic shock, vasculitis such as inflammations consequent on thrombo-phlebitis, ulcerative colitis.
• cytokines are known to be strong pro-inflammatory agents
• a series of compounds have been prepared and tested for their activity in vitro and in vivo on the synthesis of cytokines such as TNF, as well as for their action on other cellular phenomena, such as platelet aggregation and the production of free radicals, as well as their action in in vivo models of inflammation.
• the non-steroid compounds are known for their poor, or lack of, ability to inhibit the release of inflammatory cytokines, that are, on the contrary, inhibited by steroid-type compounds. Since these compounds have various toxic activities, the availability of non-steroid products specifically active on cytokine synthesis is particularly useful in developing innovative therapies.
• Preferred compounds of the present invention are the following:
• Test 1 Inhibition of TNF Release After Stimulation with LPS in vitro
• test compound was added to the culture medium in a murine macrophage line (J774) or to whole blood anti-coagulated with heparin.
• the cells were then stimulated with bacterial lipopolysacccharide. After incubation at 37° C. for a suitable length of time, the supernatant was removed from the culture and incubated with a line of murine fibroblasts (L929), sensitive to TNF.
• L929 murine fibroblasts
• the quantity of TNF released after stimulation with LPS is measured by comparing mortality of the L929 cells with that of the controls. Table 1 reports for the single compounds the concentration ( ⁇ standard error) able to inhibit fibroblast mortality (L929) by 50%.
• Test 2 Inhibition of the Release of TNF and IL- ⁇ Following Stimulation with LPS in vivo
• Conscious rats were injected with a dose of LPS, which is able to stimulate the release of TNF and IL-1 ⁇ , having first received an intravenous administration (15 minutes beforehand) of the test compound at a dose of 0.5 mg/Kg or 2 mg/Kg. Blood levels of TNF and IL-1 ⁇ were measured by the ELISA method on blood samples taken 75 minutes and 120 minutes after LPS respectively.
• Table 2 below reports the percent of inhibition of each product compared to that of the controls treated with saline.
• TABLE 2 Dose of 0.5 mg/Kg Dose of 2 mg/Kg % of % of % of % of inhibition of inhibition of inhibition of inhibition of inhibition of Compound TNF IL-1 ⁇ TNF IL-1 ⁇ 201006 9 45 30 28 (Proendotel) 203152 72 64 73 60 201273 72 67 84 39 201326 85 40 77 62 201330 75 33 62 57
• Test 3 Reduction in Inflammation in Carrageenin-Induced Oedema in Rat Paw
• a model of acute inflammation induced by intraplantar injection of 1.5 mg of carrageenin in rat paw was used.
• Administration of the compounds (2 mg/Kg) was by the intravenous route 5 minutes before oedema was induced: 3 hours later the animals were sacrificed and their paws were weighed as an indication of inflammation.
• the control animals received saline instead of the test compounds.
• the weight of the paws of the control animals was taken to correspond to 100% on the inflammation index.
• Test 4 Inhibition of the Release of Nitrite-Nitrate in Rat Plasma
• Test 5 Inhibition of the Formation of Superoxide anions Induced by f-MLP in Human Whole Blood
• Test 6 Inhibition of Collagen-Induced Platelet Aggregation in Human Whole Blood
• the aggregation event was analysed by counting the single non-aggregated platelets with a cell counter 5 minutes after adding collagen (1 ⁇ g/ml). In these working conditions, collagen induces platelet aggregation of over 80%. The test compounds or saline as control were preincubated for 1 minute before the collagen was added.
• Table 6 reports the concentrations that inhibit platelet aggregation by 50% compared to saline. TABLE 6 Compound CI 50 ⁇ M/l PG12 0.05 201006 (Proendotel) 35.0 203152 >100 201261 >100 201273 >100 201326 78 201330 93
• Test 7 Toxicity After a Single Intravenous Administration
• mice Male CD-1 mice were used to assess the maximum non-lethal does (MNLD—the highest does at which no cases of mortality are observed) and the maximum tolerated does (MTD—does at which no evident or marked signs of tocicity or altered behaviour are observed). Results are shown in Table 7. TABLE 7 Compound DMT (mg/Kg) DMNL (mg(Kg) 201006 (Proendotel) 6.25 12.5 203152 50 200 201261 50 200 201273 25 100 201326 6.25 50 201330 25 50
• Test 8 Metagenesis (Mini-Ames Test)
• Mini-Ames test is a version of the traditional Ames test which involves the incubation of bacterial cells on 35-mm dishes instead of 100-mm dishes, and it requires a smaller quantity of product while maintaining the reliability of the result.
• the test is conducted on two strains of Salmonella typhimurium requiring histidine (TA 98 and TA 100) and two strains of Escherichia coli requiring tryptophan (WP2 pKM101 and WP2 uvrA pKM1091).
• the bacterial cells were exposed to different concentrations of the test compounds in the presence and absence of a microsomal liver enzyme preparation, to reveal any possible metabolite activity.
• Mutagenic activity was determined as the ability of the test compound to induce a significant increase in the number of mutant clones compared to those developed spontaneously in the cultures with the control vehicle.
• the compounds of the invention can be prepared by known methods (Claisen's condensation and rearrangement, Pechmann's condensation, Williamson's synthesis, Fischer's esterification).
• the coumarins are synthesised by condensation between a resorcin and a ⁇ -ketoester. They can then be alkylated by substituting the aromatic hydrogens by reaction with formaldehyde and an amine.
• the hydroxyl can be alkylated with an alkylating agent or with a spacer (an alkyl dihalide or an alkyl halide substituted with an epoxy ring, an ester, an amine) that may then be further alkylated.
• the synthesised product may undergo hydrogenation to reduce the double bond between positions 3 and 4, or the oxygen in position 2 can be substituted by reaction with Lawesson's reagent.
• reaction lasts 8 hours, after which the solvent is evaporated, and the residue redissolved in ethyl acetate, washed with 1N NaOH, crystallised from ethanol and vacuum-dried.
• the resulting product is vacuum-heated to 200° C. till the accumulated heat indicates Claisen's transposition reaction. Once cooled, the mass is redissolved in ethyl acetate and the transposed product is extracted with 1N NaOH (4-methyl-7-hydroxy 6 or 8 allyl hymecromone) which is then precipitated with 1N HCl.
• the aqueous solution is neutralised with NaHCO 3 , extracted with methylene chloride and purified by silica gel chromatography with methylene chloride-methanol-ammonia 30% at a gradient of between 98-2-0.2 and 90-10-0.2.
• the clean fractions are concentrated, hydrolysed in 100 ml of 1N NaOH/tert-butanol 1:1, and neutralised with HCl.
• the pH is adjusted to 8 with ammonia and the mixture is extracted with chloroform/n-butanol 1:1.
• the residue is purified by silica gel chromatography with eluent CH2Cl2-CH3OH-ammonia 30%, at a gradient of between 98-2-0.2 and 90-5-0.4.
• the clean fractions are concentrated to dryness, redissolved in ethyl acetate and treated with HCl in ethanol until the Congo red indicator has changed colour, then concentrated and freeze-dried from water (yield 13.5 g, M.W. 431.96).

Landscapes

• Health & Medical Sciences (AREA)
• Chemical & Material Sciences (AREA)
• Organic Chemistry (AREA)
• General Health & Medical Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Veterinary Medicine (AREA)
• Public Health (AREA)
• Animal Behavior & Ethology (AREA)
• Engineering & Computer Science (AREA)
• Pharmacology & Pharmacy (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Medicinal Chemistry (AREA)
• General Chemical & Material Sciences (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Cardiology (AREA)
• Diabetes (AREA)
• Hematology (AREA)
• Heart & Thoracic Surgery (AREA)
• Molecular Biology (AREA)
• Genetics & Genomics (AREA)
• Biochemistry (AREA)
• Biotechnology (AREA)
• Immunology (AREA)
• Communicable Diseases (AREA)
• Oncology (AREA)
• Obesity (AREA)
• Pulmonology (AREA)
• Pain & Pain Management (AREA)
• Toxicology (AREA)
• Dermatology (AREA)
• Rheumatology (AREA)
• Urology & Nephrology (AREA)
• Vascular Medicine (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
• Pyrane Compounds (AREA)
• Nitrogen And Oxygen Or Sulfur-Condensed Heterocyclic Ring Systems (AREA)
• Medicinal Preparation (AREA)
• Saccharide Compounds (AREA)

Applications Claiming Priority (3)

Publications (1)

Family

ID=11452041

Family Applications (1)

Country Status (9)

Cited By (1)

Families Citing this family (4)

Citations (2)

Family Cites Families (6)

• 2000
  • 2000-07-31 IT IT2000PD000193A patent/IT1317356B1/it active
• 2001
  • 2001-07-26 US US10/343,241 patent/US20040023893A1/en not_active Abandoned
  • 2001-07-26 ES ES01962875T patent/ES2247158T3/es not_active Expired - Lifetime
  • 2001-07-26 CA CA002417788A patent/CA2417788A1/fr not_active Abandoned
  • 2001-07-26 AU AU8395901A patent/AU8395901A/xx active Pending
  • 2001-07-26 DE DE60113278T patent/DE60113278T2/de not_active Expired - Fee Related
  • 2001-07-26 WO PCT/EP2001/008642 patent/WO2002010148A1/fr active IP Right Grant
  • 2001-07-26 EP EP01962875A patent/EP1307438B1/fr not_active Expired - Lifetime
  • 2001-07-26 AU AU2001283959A patent/AU2001283959B8/en not_active Ceased
  • 2001-07-26 JP JP2002515879A patent/JP2004505072A/ja active Pending

Patent Citations (2)

Also Published As

Similar Documents

Legal Events